EP0395137B1 - Sensor with antigen chemically bonded to a semiconductor device - Google Patents

Sensor with antigen chemically bonded to a semiconductor device Download PDF

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Publication number
EP0395137B1
EP0395137B1 EP90200915A EP90200915A EP0395137B1 EP 0395137 B1 EP0395137 B1 EP 0395137B1 EP 90200915 A EP90200915 A EP 90200915A EP 90200915 A EP90200915 A EP 90200915A EP 0395137 B1 EP0395137 B1 EP 0395137B1
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EP
European Patent Office
Prior art keywords
antigen
eos
silicon oxide
immunochemical
protein
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Lifetime
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EP90200915A
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German (de)
English (en)
French (fr)
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EP0395137A3 (en
EP0395137A2 (en
Inventor
Claudio Colapicchioni
Filippo Porcelli
Carlo Antonio Nuzzolo
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Eni Tecnologie SpA
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Eniricerche SpA
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
    • C07F7/02—Silicon compounds
    • C07F7/08—Compounds having one or more C—Si linkages
    • C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
    • C07F7/1804—Compounds having Si-O-C linkages
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/403—Cells and electrode assemblies
    • G01N27/414—Ion-sensitive or chemical field-effect transistors, i.e. ISFETS or CHEMFETS
    • G01N27/4145—Ion-sensitive or chemical field-effect transistors, i.e. ISFETS or CHEMFETS specially adapted for biomolecules, e.g. gate electrode with immobilised receptors
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • This invention relates to a semiconductor device of EOS (electrolyte oxide semiconductor) or CHEMFET (chemical field effect transistor) type containing a polysiloxane matrix and an immunochemical membrane formed from a monolayer consisting of an antigen or a polymeric multilayer consisting of an antigen and a protein.
  • EOS electronic oxide semiconductor
  • CHEMFET chemical field effect transistor
  • the hematic concentration of metabolytes, hormones or proteins is determined using various processes employing immunofluorescent (FIA etc.), immunoradiometric (RIA, IRMA etc.) or immunoenzymatic (ELISA etc.) systems.
  • FAA immunofluorescent
  • RIA immunoradiometric
  • IRMA IRMA
  • ELISA immunoenzymatic
  • the resultant immunochemical membrane is rather unstable and in particular is subject to interference by other chemical species such as ions or proteins present in solution.
  • EP-A-0 291 130 discloses the preparation of enzymatic membranes bound to a surface.
  • EP-A-0 127 438 discloses only generically the possibility of using a device for measuring a physical characteristic for binding an antigen through a chemical bond or an affinity type bond, whereas no example or data are disclosed with regard to the antigen on FET or on other devices.
  • an immunochemical membrane consisting of a functionalized antigen (and possibly a protein) chemically bonded to the surface silicon oxide of the device by a polysiloxane matrix, the membrane maintains its biological functionality constant for more than one month if stored in a pH 7.8 buffered solution.
  • interference problems can be overcome by marking the antibody directed against the antigen to be determined, or marking a second antibody directed against this antibody, with an enzyme such as glucose oxidase or urease, which during the catalyzed reaction produces electrical charges detected by the transducer (EOS or CHEMFET).
  • an enzyme such as glucose oxidase or urease
  • AEAPS aminoethylaminopropyltrimethoxysilane
  • AAPS anthranylamidepropyltriethoxysilane
  • an EOS device Besides containing surface silicon oxide an EOS device must contain in the part below the silicon oxide a layer of aluminium or gold deposited by evaporation.
  • a functional organosilane of formula (2) its preparation process comprises reacting isatoic anhydride or its derivatives with amino silanes of type in accordance with the following scheme:
  • the preferred silanes for preparing the functional organosilanes for the purposes of the present invention are: 3-aminopropyltriethoxysilane H2N-(CH2)3-Si-(OC2H5)3 aminomethyltriethoxysilane H2N-CH2-Si-(OC2H5)3 2-aminoethyl-aminopropyltrimethoxysilane H2N-(CH2)2-NH-(CH2)3-Si-(OCH3)3 2-aminoethyl-aminopropyltriethoxysilane H2N-(CH2)2-NH-(CH2)3-Si-(OC2H5)3 2-aminoethyl-aminopropylmethyldimethoxysilane
  • the synthesis reaction is conducted by gradually adding the isatoic anhydride to the aminosilane in the presence or absence of solvent.
  • reaction is carried out in solvent this must not react with the isatoic anhydride.
  • mild reaction conditions must be used, for example, with no catalysts.
  • Alcohols, ethers, chlorinated solvents etc. can be used as solvents.
  • the process must be conducted in the absence of water, which would hydrolyze the alkoxy groups.
  • the reaction can also be effected by adding the isatoic anhydride in one addition taking account of the slight exothermic nature of the reaction (cooling is required) and the fact that in spite of the considerable reactivity difference between the aliphatic amine (aminosilane) and the aromatic amine (reaction product), oligomer products of the type could form, even if only in minimum quantity.
  • Enzyme types which produce electrical charges during the catalyzed reaction can be introduced into said device.
  • Any proteins used can be chosen for example from albumin, hemocyanin etc.
  • the said immunosensor can be used to determine, in biological fluids, particular circulating antibodies such as those directed against malaria plasmodium or the AIDS virus (HTLVIII), the detection of which is important in clinical diagnostics.
  • circulating antibodies such as those directed against malaria plasmodium or the AIDS virus (HTLVIII)
  • HTLVIII AIDS virus
  • An alternative measurement process is to use a second antibody directed towards the first, or protein A or protein G of microbic origin, marked with an enzyme such as glucose oxidase or urease or others having the aforesaid characteristics.
  • the immunosensor of the present invention can be used to determine contaminant substances such as atrazine herbicides, paraquat, molinate or others for which the relative functionalized antigen can be constructed.
  • Atrazine is a herbicide widely used in agriculture mainly for maize cultures, and acts by interfering with the photosynthesis system of the infesting plants.
  • the selectivity with respect to maize is due to the fact that the active principle is degraded by the culture by means of specific enzymes or polyphenols, into a non-toxic substance (hydroxyatrazine).
  • this herbicide can cause pollution of waters surrounding the treated cultures, where it can remain for a long time before undergoing natural degradation.
  • the determination of atrazine in aqueous solutions is commonly effected by complex analytical processes such as gas chromatography (GC), HPLC or GLC after a preliminary purification procedure.
  • GC gas chromatography
  • HPLC HPLC
  • GLC GLC after a preliminary purification procedure.
  • the use of the immunosensor of the present invention wold have the advantage over the aforesaid processes of being able to directly analyze the samples in aqueous solution.
  • an atrazine analogue namely ametryne
  • ametryne is used functionalized as ametryne sulphoxide and bonded by this reactive group to a carrier (generally a protein such as albumin, hemocyanin or others).
  • This conjugate once inoculated into a rabbit (or another suitable experimental animal) induces the production of antibodies which recognize both ametryne and atrazine itself.
  • antibodies can be obtained against other contaminant substances such as paraquat, molinate etc., as described in the literature [eg. J. Van Emon, B. Hammock. J.N. Seiber; Anal. Chem. (1986), 58, 1866-1873; S.J. Gee, T. Miyamoto, M.H. Goodrow, D. Buster, B.D. Hammock; J. Agric. Food Chem. (1988), 36, 863-870].
  • ametryne-protein conjugate can be chemically bonded to the device by bifunctional coupling reactants, or ametryne sulphoxide can be directly bonded thereto by said reactive group.
  • the present invention also provides processes which can be used to form the device with an immunochemical membrane.
  • a first process for constructing the sensor with the immunochemical membrane formed from a monolayer consisting of a functionalized antigen comprises the following steps:
  • a second process for constructing the sensor with the immunochemical membrane formed from a polymer multilayer consisting of a functionalized antigen and a protein comprises the following steps:
  • the bifunctional coupling agents used in the aforesaid process can be chosen from dialdehydes (such as glutaraldehyde) or from diisocyanates (such as toluene 2.4-diisocyanate).
  • Deposition of the siloxane prepolymer in the aforesaid processes can be accomplished by spin-on techniques, and the polysiloxane layer can be activated and reacted with the antigen and protein on a rocking plate.
  • the thermal curing of the siloxane prepolymer is effected at a temperature of between 80 and 140°C, the subsequent immunochemical immobilization treatment being between 25 and 37°C.
  • the thickness of the deposited siloxane prepolymer must be between 0.5 and 3 »m, and the thickness of the immunochemical membrane can be between 0.5 and 2 »m.
  • the rotary disc apparatus To form the polysiloxane layer the rotary disc apparatus must operate at a speed preferably of between 400 and 4500 rpm during the depositions.
  • the siloxane prepolymer can also be deposited on silicon or silicon oxide substrates of EOS or ISFET devices by plasma deposition, preferably under the following conditions:
  • the preferred silanes are AEAPS and 3-APTS.
  • the present invention also provides a process for measuring the unknown antigen concentration using the EOS or CHEMFET electronic devices as transducers.
  • Determination of the antigen concentration is accomplished by marking the antibody directed against the antigen itself, or marking a second antibody directed against the antigen-recognizing antibody, with an enzyme such as glucose oxidase, urease or others, which during the catalyzed reaction produces electrical charges detected by the transducer (EOS or FET).
  • an enzyme such as glucose oxidase, urease or others, which during the catalyzed reaction produces electrical charges detected by the transducer (EOS or FET).
  • the antibody-enzyme conjugate is made to compete with the antigen to be determined.
  • the quantity of marked antibody bonded to the immunochemical membrane present on the device can be determined by the electrical response of the enzyme in the presence of its substrate. This quantity is therefore inversely proportional to the quantity of antigen present in solution to be determined.
  • concentration of the antigen to be determined is again inversely proportional to the quantity of marked antibody bonded to the first antibody, determined on the basis of the enzyme electrical response recorded by the transducer.
  • This process has therefore the advantage of less interference due to ions or to other proteins present in the sample under examination, as the measurement is obtained by the electrical response of the enzyme, which marks an antibody directed against the antigen or a second antibody directed against the first antibody, when in the presence of its substrate.
  • Ametryne sulphoxide is prepared from commercial ametryne as reported by Huber S.J. (1985) in Chemosphere vol. 14 No. 11/12, pp. 1795-1803.
  • the horizontal axis represents wavelength in cm ⁇ 1 and the vertical axis represents percentage transmittance).
  • An immunosensor was formed enabling an immunochemical membrane to be obtained from a monolayer consisting of a functionalized antigen (ametryne sulphoxide) bonded chemically to the polysiloxane matrix.
  • AEAPS amino-ethylaminopropyltrimethoxysilane
  • Three depositions were effected by a rotary disc apparatus rotating at 3500 rpm for 20 seconds.
  • the mixture was incubated for 5 days in the dark at 37°C,dialyzed against 5 litres of 0.9% NaCl and lyophilized.
  • the lyophilized product was used to produce antibodies as shown in Table 1.
  • Figure 2 shows the absorption spectra of BSA (a), commercial ametryne (b) and the compound BSA-ametryne (c). (The horizontal axis represents wavelength in »m and the vertical axis the optical density).
  • the plate was then washed 5 times with 200 »l per well of pH 7.8 25 mM TBS containing 0.05% of Tween 20.
  • the plate was then washed 5 times with 200 »l per well of pH 7.8 25 mM TBS (+ 0.05% of Tween 20).
  • the plate was then washed 5 times with 200 »l per well of pH 7.8 25 mM TBS (+ 0.05% of Tween 20).
  • the plate was then washed 5 times with 200 »l per well of pH 7.8 25 mM TBS (+ 0.05% of Tween 20).
  • the substrate consisted of: 9 ml of pH 5.0 0.1 M acetate buffer and 1 ml of a 3,3′,5,5′tetramethylbenzidine solution (1 mg/ml in 0.1 M citric acid + 1.5 »l of 35% hydrogen peroxide).
  • Figure 3 shows the curves representing titration of anti-ametryne antiserum with BSA-ametryne (*) and hemocyanin-ametryne (#) supported in the plate.
  • EOS-ametryne and other EOS (AEAPS) samples used as blanks were incubated with 0.5 ml of immune serum diluted to between 1:500 and 1:4000 in pH 7.8 50 mM TBS (+ 0.5% casein + 0.1% Triton X100 + 2% PEG 6000) for 30 minutes at ambient temperature.
  • TMB substrate
  • EOS EOS
  • EOS-ametryne were incubated with 0.5 ml of immune serum diluted to 1:2000 + atrazine (0.05, 1, 5 and 25 »g/l) in pH 7.8 50 mM TBS (+ 0.5% casein + 0.1% Triton X100 + 2% PEG 6000) for 30 minutes at ambient temperature.
  • the chip (HEDCO, University of Utah), having a size of 1.28 mm x 2.16 mm and comprising two 400 »m x 20 »m gates, was stuck onto a slide by adhesive tape (3M electrical tape 92) to protect the electrical contact region and edges but leaving the gates free.
  • a layer of AEAPS [3(2-aminoethyl)aminopropyltrimethoxysilane] was then deposited on it as described heretofore. After depositing the polysiloxane, the chip was mounted on a type TO-5 support, contacted with an ultrasonic welder (Kulicke and Soffa mod. 4123) and then encapsulated with Epotek H-77 epoxy resin.
  • FET devices were reacted for 5 days at ambient temperature under agitation, each with 0.5 ml of a solution of about 1 mg/ml of ametryne sulphoxide in 50 mM pH 9.6 carbonate buffer, containing 10% of absolute ethanol. They were thin washed three times with 50 mM pH 7.8 TBS buffer containing 10% of ethanol and then twice with the same buffer without ethanol.
  • the devices were incubated with 0.5 ml of immune serum diluted 1:500 in 50 mM pH 7.8 TBS buffer containing 2.5% BSA, 0.1% Triton X-100 and 2% PEG 6000 (Incubation Buffer), for 30 minutes at ambient temperature.
  • Figure 6 shows the curve of immunosensor electrical response after the addition of 0.05 M glucose (at time a) and after the addition of the Wash Buffer and 20 mM pH 7.0 phosphate (at time b).
  • the horizontal axis represents time in minutes and the vertical axis represents the electrical response in mV.
  • FETs were reacted for 1 hour at ambient temperature under agitation with 0.250 ml of a 10 mg/ml solution of BSA-ametryne in 20 mM pH 7.0 phosphate buffer, in the presence of 0.1% glutaraldehyde, and were then washed with only buffer.
  • the chip was incubated for about 10 minutes with 0.1 M pH 2.0 glycine-HCl buffer to obtain antigen-antibody separation, so regenerating the device to allow it to be used for a further measurement.
  • Figure 7 shows the response in mV (vertical axis) obtained during various cycles effected with the same device.
  • the FET-BSA-ametryne was incubated with 0.5 ml of immune serum diluted 1:500 in Incubation Buffer for 30 minutes at ambient temperature, and washed 4 times with the Wash Buffer.

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  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Organic Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)
  • Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
  • Bipolar Transistors (AREA)
  • Saccharide Compounds (AREA)
EP90200915A 1989-04-21 1990-04-13 Sensor with antigen chemically bonded to a semiconductor device Expired - Lifetime EP0395137B1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IT8920259A IT1229691B (it) 1989-04-21 1989-04-21 Sensore con antigene legato chimicamente a un dispositivo semiconduttore.
IT2025989 1989-04-21

Publications (3)

Publication Number Publication Date
EP0395137A2 EP0395137A2 (en) 1990-10-31
EP0395137A3 EP0395137A3 (en) 1991-06-12
EP0395137B1 true EP0395137B1 (en) 1995-08-16

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EP90200915A Expired - Lifetime EP0395137B1 (en) 1989-04-21 1990-04-13 Sensor with antigen chemically bonded to a semiconductor device

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US (1) US5160597A (it)
EP (1) EP0395137B1 (it)
JP (1) JPH02296142A (it)
AT (1) ATE126597T1 (it)
DE (1) DE69021621T2 (it)
DK (1) DK0395137T3 (it)
ES (1) ES2075135T3 (it)
GR (1) GR3017224T3 (it)
IT (1) IT1229691B (it)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6060327A (en) * 1997-05-14 2000-05-09 Keensense, Inc. Molecular wire injection sensors
US6699667B2 (en) 1997-05-14 2004-03-02 Keensense, Inc. Molecular wire injection sensors

Families Citing this family (31)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU3809993A (en) * 1992-03-16 1993-10-21 Immuna Care Corporation Improved test kit for the determination of steroids and related compounds
CA2108705A1 (en) * 1992-11-06 1994-05-07 Richard Barner Biologically recognizing layers on new ti02 waveguide for biosensors
FR2722294B1 (fr) * 1994-07-07 1996-10-04 Lyon Ecole Centrale Procede d'analyse qualitative et/ou quantitative de substances biologiques presentes dans un milieu liquide conducteur et capteurs biochimiques d'affinite utilises pour la mise en oeuvre de ce procede
US7468398B2 (en) 1994-09-06 2008-12-23 Ciba Vision Corporation Extended wear ophthalmic lens
US5760100B1 (en) 1994-09-06 2000-11-14 Ciba Vision Corp Extended wear ophthalmic lens
US6355436B1 (en) 1996-05-17 2002-03-12 L'ecole Centrale De Lyon Method for analyzing biological substances in a conductive liquid medium
SE9602545L (sv) * 1996-06-25 1997-12-26 Michael Mecklenburg Metod för att diskriminera komplexa biologiska prover
EP1060395B1 (en) 1998-02-04 2008-04-30 Invitrogen Corporation Microarrays and uses therefor
US6897073B2 (en) 1998-07-14 2005-05-24 Zyomyx, Inc. Non-specific binding resistant protein arrays and methods for making the same
US6682942B1 (en) 1998-07-14 2004-01-27 Zyomyx, Inc. Microdevices for screening biomolecules
US6406921B1 (en) * 1998-07-14 2002-06-18 Zyomyx, Incorporated Protein arrays for high-throughput screening
US20020119579A1 (en) * 1998-07-14 2002-08-29 Peter Wagner Arrays devices and methods of use thereof
US6576478B1 (en) 1998-07-14 2003-06-10 Zyomyx, Inc. Microdevices for high-throughput screening of biomolecules
US7371563B2 (en) * 2000-11-08 2008-05-13 Surface Logix, Inc. Peelable and resealable devices for biochemical assays
US7001740B2 (en) * 2000-11-08 2006-02-21 Surface Logix, Inc. Methods of arraying biological materials using peelable and resealable devices
US7439056B2 (en) 2000-11-08 2008-10-21 Surface Logix Inc. Peelable and resealable devices for arraying materials
US7351575B2 (en) * 2000-11-08 2008-04-01 Surface Logix, Inc. Methods for processing biological materials using peelable and resealable devices
US6967074B2 (en) * 2000-11-08 2005-11-22 Surface Logix, Inc. Methods of detecting immobilized biomolecules
US6803205B2 (en) * 2000-11-08 2004-10-12 Surface Logix, Inc. Methods of measuring enzyme activity using peelable and resealable devices
GB0105831D0 (en) * 2001-03-09 2001-04-25 Toumaz Technology Ltd Method for dna sequencing utilising enzyme linked field effect transistors
US20050053949A1 (en) * 2003-09-08 2005-03-10 Silin Vitalii Ivanovich Biochip for proteomics applications
US7318887B2 (en) * 2003-10-08 2008-01-15 Honeywell International Inc. Solid state reference electrode
US8198658B2 (en) * 2007-06-13 2012-06-12 Samsung Electronics Co., Ltd. Device and method for detecting biomolecules using adsorptive medium and field effect transistor
KR101465961B1 (ko) * 2007-10-09 2014-12-01 삼성전자주식회사 유전자 검출 방법 및 장치
US9029132B2 (en) * 2009-08-06 2015-05-12 International Business Machines Corporation Sensor for biomolecules
US8052931B2 (en) * 2010-01-04 2011-11-08 International Business Machines Corporation Ultra low-power CMOS based bio-sensor circuit
US9068935B2 (en) 2010-04-08 2015-06-30 International Business Machines Corporation Dual FET sensor for sensing biomolecules and charged ions in an electrolyte
EP2469275B1 (en) 2010-12-24 2015-12-23 Honeywell Romania S.R.L. Cantilevered carbon dioxide sensor
CA2862468A1 (en) 2012-01-23 2013-08-01 The Ohio State University Devices and methods for the rapid and accurate detection of analytes
IN2012DE00264A (it) * 2012-01-31 2015-07-10 Council Scient Ind Res
US20230236147A1 (en) * 2020-05-27 2023-07-27 Jose Maria Piccinini Sensors and systems based on field-effect transistors, methods of preparation and devices for their operation

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5825221B2 (ja) * 1977-12-12 1983-05-26 株式会社クラレ Ｆｅｔ比較電極
GB8314523D0 (en) * 1983-05-25 1983-06-29 Lowe C R Diagnostic device
GB8406955D0 (en) * 1984-03-16 1984-04-18 Serono Diagnostics Ltd Assay
US4708947A (en) * 1985-01-10 1987-11-24 Kuraray Co., Ltd. Water resistant composition and heat-sensitive recording sheet containing the same
JPH0721478B2 (ja) * 1986-03-31 1995-03-08 財団法人化学及血清療法研究所 免疫センサ−用作用膜
IT1215491B (it) * 1987-05-15 1990-02-14 Enricerche Spa Biosensore con membrana enzimatica legata chimicamente a un dispositivo semiconduttore.
US5001048A (en) * 1987-06-05 1991-03-19 Aurthur D. Little, Inc. Electrical biosensor containing a biological receptor immobilized and stabilized in a protein film

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
BIOLOGICAL ABSTRACTS, vol. 81, no. 9, 1986, Philadelphia, PA (US); S.J. HUBER, p. AB-945, no. 86703/ *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6060327A (en) * 1997-05-14 2000-05-09 Keensense, Inc. Molecular wire injection sensors
US6699667B2 (en) 1997-05-14 2004-03-02 Keensense, Inc. Molecular wire injection sensors

Also Published As

Publication number Publication date
JPH02296142A (ja) 1990-12-06
EP0395137A3 (en) 1991-06-12
IT8920259A0 (it) 1989-04-21
DK0395137T3 (da) 1995-11-06
DE69021621T2 (de) 1996-03-14
US5160597A (en) 1992-11-03
ATE126597T1 (de) 1995-09-15
EP0395137A2 (en) 1990-10-31
IT1229691B (it) 1991-09-06
ES2075135T3 (es) 1995-10-01
GR3017224T3 (en) 1995-11-30
DE69021621D1 (de) 1995-09-21

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